Targeted Gene Therapy for Brain Tumor

脑肿瘤靶向基因治疗

• 批准号: 10470298
• 负责人: HAMADA Hirofumi
• 金额: $ 8.83万
• 依托单位: Sapporo Medical University (1999)Japanese Foundation For Cancer Research (1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470298/
• 关键词: Adenoviral Vector; Cancer Gene Therapy; Fiber mutant; Apoptosis; Brain Tumor; 遺伝子治療; アデノウイルス ファイバー; EIA; p53; Bax

Highly augmented cytopathic effect of a fiber-mutant E1B-defective adenovirus for gene therapy of glioma. Cancer Res., 1999. An E1B 55-kDa gene-defective adenovirus(Adv), ONYX-015, has been reported to be a highly useful replication-competent Adv that shows cytopathic effect for cancers with an abnormal p53 gene, without damaging normal tissues. In this study, we combined this Adv (Adv-E1Adb) with a fiber mutation, F/K20, which has a stretch of 20 lysine residues added at the COO-H-terminus of the fiber and shows high transduction efficiency to gliomas. In U-373 MG glioma cells, the transduction efficiency of Adv-F/K20 for lacZ was nine times higher than that of the Adv with wild-type fiber (Adv-F/wt) for lacZ. At a multiplicity of infection of 30, the replication efficiency of Adv-E1AdB-F/K20 was 11 times higher than that of Adv-E1AdB with wt fiber (Adv-E1AdB-F/wt). The ED50 value of AdvE1AdB-F/K20 to U-373 MG cells, which is a measure of the in vitro cytopathic effect, was 32 times g … More reater than that of Adv-E1AdB-F/wt. Injection of Adv-E1AdB-F/K20 suppressed the in vivo growth of tumors. The antitumoral effect of Adv-E1AdB-F/K20 was remarkably stronger than that of Adv-E1AdB-F/wt. A greater quantity of replicated virus protein (hexon) by infection with Adv-E1AdB-F/K20 was demonstrated in vitro and in vivo, compared with that of Adv-E1AdB-F/wt. In conclusion, gene therapy using Adv-E1AdB-F/K20, which drastically augmented the antitumoral effect of Adv-E1AdB, will be promising therapeutic approach for gliomas.Adenovirus-mediated transfer of p33ィイD1INGィエD1 with p53 drastically augments apoptosis in gliomas. Cancer Res., 1999. The p53 tumor suppressor gene is an important target for the gene therapy of cancers, and clinical trials targeting this gene have been conducted. Some cancers, however, are refractory to p53 gene therapy. Therefor, it has been combined with other therapies including chemotherapy and radiotherapy to enhance the cytopathic effect of p53 induction. The p33ィイD1ING1ィエD1 gene cooperates with p53 to block cell proliferation. In this study, we investigated whether adenovirus (Adv)-mediated co-induction of p33ィイD1ING1ィエD1 and p53 enhances apoptosis in glioma cells (U251 and U-373MG), which showed no genetic alterations but low expression levels of p33ィイD1ING1ィエD1. Co-infection of Adv-p33 and Adv-MBP-p53 at the same MOIs induced drastically enhanced apoptosis in both cell lines. Our results indicated that this co-infection approach can be used as a modality for the gene therapy of gliomas, sparing damage to normal tissues. Less

神经胶质瘤基因治疗中纤维突变E1 B缺陷腺病毒的细胞病变效应增强癌症研究所，1999.据报道，E1 B 55-kDa基因缺陷型腺病毒（Adv）ONYX-015是高度有用的具有复制能力的Adv，其对具有异常p53基因的癌症显示细胞病变效应，而不损害正常组织。在这项研究中，我们将这种Adv（Adv-E1 Adb）与纤维突变F/K20相结合，F/K20在纤维的COO-H-末端添加了一段20个赖氨酸残基，并显示出对胶质瘤的高转导效率。在U-373 MG胶质瘤细胞中，Adv-F/K20对lacZ的转导效率比具有野生型纤维的Adv（Adv-F/wt）对lacZ的转导效率高9倍。在感染复数为30时，Adv-E1 AdB-F/K20的复制效率比含有wt纤维的Adv-E1 AdB（Adv-E1 AdB-F/wt）高11倍。AdvE 1AdB-F/K20对U-373 MG细胞的ED 50值（其是体外细胞病变效应的量度）为32倍g ...更多信息 比Adv-E1AdB-F/wt.注射Adv-E1 AdB-F/K20抑制肿瘤的体内生长。Adv-E1 AdB-F/K20的抗肿瘤作用明显强于Adv-E1 AdB-F/wt。在体外和体内实验中，与Adv-E1 AdB-F/wt相比，Adv-E1 AdB-F/K20感染的病毒蛋白（六邻体）复制量更大。结论：腺病毒介导的p33介导的p53介导的p53介导的p33-D1 ING-D1基因转染可显著促进胶质瘤细胞的凋亡，腺病毒介导的p53介导的p33-D1 ING-D1基因转染可显著促进胶质瘤细胞的凋亡。癌症研究所，1999. p53肿瘤抑制基因是癌症基因治疗的重要靶点，已经进行了针对该基因的临床试验。然而，一些癌症对p53基因治疗是难治的。因此，它已与其他疗法包括化疗和放疗相结合，以增强p53诱导的细胞病变效应。p33启动子D1 ING 1启动子D1基因与p53协同作用阻断细胞增殖。在这项研究中，我们研究了腺病毒（Adv）介导的p33 β D1 ING 1 β D1和p53的共诱导是否会增强胶质瘤细胞（U251和U-373 MG）的凋亡，这表明没有遗传改变，但p33 β D1 ING 1 β D1的低表达水平。以相同MOI共感染Adv-p33和Adv-MBP-p53在两种细胞系中诱导急剧增强的凋亡。我们的研究结果表明，这种联合感染的方法可以作为一种方式的基因治疗的胶质瘤，不损害正常组织。少

项目成果

Yoshida,Y., Sadata,A., Zhang,W., Shinoura,N., and Hamada,H.: "Generation of fiber-mutant recombinant adenoviruses for gene therapy of malignant glioma." Human Gene Therapy. 9(17). 2503-2515 (1998)

Yoshida,Y.、Sadata,A.、Zhang,W.、Shioura,N. 和 Hamada,H.：“用于恶性神经胶质瘤基因治疗的纤维突变重组腺病毒的生成。”

Zhang,W., He L., Yuan Z., Xie Z., Wang,J., Hamada,H., and Cao X.: "Enhanced therapeutic efficacy of tumor RNA-pulsed dendritic cells after-genetic modification with lymphotactin." Human Gene Therapy. 10(7)in press. (1999)

张 W.、何 L.、袁 Z.、谢 Z.、王 J.、滨田 H. 和曹 X.：“用淋巴趋化素进行基因修饰后肿瘤 RNA 脉冲树突状细胞的治疗效果增强。

Motoi, F., et al. and Hamada, H.: "Effective gene therapy for pancreatic canncer by cytokines mediated by restricted replication-competent adenovirus"Hum. Gene ther.. 11(in press). (2000)

本井，F.，等人。

TANAKA,H., YOSHIZAWA,H., YAMAGUCHI,Y., ITO,K., KAGAMU,H., SUZUKI,E., HAMADA,H., ARAKAWA,M.,: "Successful adoptive immunotherapy of murine nonimmunogenic tumor with specific effector cells generated from gene-modified tumor-primed lymph node cells."J. Immu

TANAKA,H., YOSHIZAWA,H., YAMAGUCHI,Y., ITO,K., KAGAMU,H., SUZUKI,E., HAMADA,H., ARAKAWA,M.,：“小鼠非免疫原性肿瘤的成功过继免疫治疗

SHINOURA,N., YOSHIDA,Y., ASAI,A., KIRINO,T., HAMADA,H.: "Relative level of expression of Bax and Bcl-XL determines the cellular fate of apoptosis/necrosis induced by the overexpression of Bax."Oncogene. 18(41). 5703-5713 (1999)

Shinoura,N.、YOSHIDA,Y.、ASAI,A.、KIRINO,T.、HAMADA,H.：“Bax 和 Bcl-XL 的相对表达水平决定了 Bax 过表达诱导的细胞凋亡/坏死的细胞命运